FR2738745A1 - NOVEL COMPOSITIONS BASED ON A SYNERGETIC MIXTURE BETWEEN AT LEAST ONE VDR LIGAND AND A RETINOID, AND USES THEREOF - Google Patents

Abstract

A novel combination of one or more ligands with VDR receptor selective activity and one or more retinoids selective for RAR gamma receptors as opposed to RAR alpha receptors is described, as well as the use thereof in cosmetic and skin-care applications for treating disorders related to a hyperproliferation of cells, in particular skin cells, such as psoriasis.

Description

The invention relates to a new association, in particular a pharmaceutical association, comprising a synergistic association between at least one ligand having activity for nuclear receptors of VDR type and at least one particular retinoid, as well as the use of this composition in the pharmaceutical field or cosmetic.

It is known that a ligand exhibiting activity for nuclear receptors of the VDR type (vitamin D3 receptor), such as 11a, 25-dihydroxyvitamin D3 or its analogs, inhibits the proliferation of keratinocytes. So, this type of compound is used to treat psoriasis. However, these compounds, such as 1a, 25-dihydroxyvitamin D3, have side effects, hypercalcemia or hypercalciuria, due to an increase in serum calcium levels.

We know that retinoic acid is a modulator (i.e. an inhibitor or, on the contrary, a stimulator, depending on the nature of the cells treated) of the differentiation and / or of the proliferation of many normal or transformed cell types. For example, it inhibits the differentiation of epithelial cells, such as keratinocytes of the epidermis, without being particularly active on the proliferation of these cells. It also inhibits the proliferation of many transformed cells, such as melanoma cells.

It is generally known that all-trans retinoic acid acts on the differentiation and / or proliferation of cells by interacting with nuclear receptors or RARs (Retinoic Acid Receptors) contained in the cell nucleus. Many synthetic structural analogues of all-trans retinoic acid or 9-cis-retinoic acid, commonly called "retinoids", have been described to date in the literature.

Some of these retinoids have already been associated with vitamin derivatives
D. Thus, in patent application AU-A-37161/93 filed by the company
Hoffmann-La Roche, 9-cis- or 13-cis-retinoic acid is associated with derivatives of vitamin D. However, these associations prove to be unsatisfactory in particular in the treatment of psoriasis. We therefore understand the need to find compositions with remarkable efficacy, especially in the treatment of psoriasis.

One of the aims of the present invention is therefore to be able to have available a composition having a remarkable inhibition of the proliferation of skin cells, and more particularly keratinocytes, allowing this composition to be used in the treatment of related dermatological disorders. to hyperproliferation of these cells.

Another object of the present invention is to be able to have a medicament which diminishes, or even suppresses, the side effects of ligands exhibiting activity for nuclear receptors of the VDR type.

The Applicant has now discovered that the association, new in itself, and in particular as a medicament, of at least one ligand having activity for nuclear receptors of VDR type and a particular retinoid which is chosen from acid 6 [3- (1 -adamantyl) -4-hydroxyphenyl] -2-naphtanoï
6- [3- (I-adamantyl) -methoxyphenyl] -2-naphthanoic acid or one of their derivatives, made it possible to inhibit, in a quite remarkable manner, the proliferation of keratinocytes. This result is all the more unexpected and surprising since these retinoids, when used alone, have no, or substantially no, specific antiproliferative activity with respect to these same cells.

This discovery is the basis of the present invention.

This association in accordance with the invention naturally finds, given the remarkable activities which it exhibits with respect to keratinocytes, a preferred application in the treatment of dermatological disorders linked to hyperproliferation of skin cells, and more particularly of keratinocytes.

Thus, in one of its first aspects, the present invention relates to a new association, in particular pharmaceutical, characterized in that it comprises, in a physiologically acceptable support, at least one ligand having activity for the VDR receptor and at least one retinoid chosen from 6 [3- (1-adamantyl) 4-hydroxyphenyl] -2-naphthanoic acid, 6- [3- (1 adam antyl) -4-m ethoxyphenyl] -2- acid naphthanoic or their derivatives.

The ligands exhibiting activity for the VDR receptor are used for the treatment of osteoporosis, it is therefore possible to use this combination according to the invention for the treatment of osteoporosis.

However, this association proves to be very advantageous for a treatment intended to inhibit the proliferation of skin cells, and more particularly of keratinocytes, in particular for the treatment of psoriasis.

A subject of the invention is therefore the use of at least one retinoid chosen from 6 [3- (1-adam antyl) 4-hydroxyphenyl] -2-naphthano ï acid that 6- [3- (1 adamantyl) 4-methoxyphenyl] -2-naphthanoic or one of their derivatives for the manufacture of a pharmaceutical composition intended to increase the activity of inhibiting proliferation of skin cells, more particularly keratinocytes, due to at least one ligand with receptor activity
VDR used for the treatment of dermatological disorders linked to hyperproliferation of skin cells.

Thus, in the context of this use according to the invention, the retinoid can be administered, simultaneously or not, by routes identical or not to the administration of the ligand having activity for the VDR receptor.

In the case where the retinoid is administered simultaneously and therefore by a route identical to the administration of the ligand having activity for the VDR receptor, the invention finally relates to the use of the above combination for the manufacture of a pharmaceutical composition, more particularly a dermatological composition, intended for treating a dermatological disorder linked to a hyperproliferation of skin cells, and more particularly of keratinocytes. This composition is more particularly intended for the treatment of psoriasis.

We understand, in skin cells, keratinocytes, melanocytes, fibroblasts, Merkel cells and Langerhans cells.

In general, it will be noted that the doses of active agents to be used to obtain the desired effect may be low, which constitutes an important advantage when it is a question of coping with problems of side effects undesirable that may appear in the organisms to be treated or during treatment, such as hypercalcemia or hypercalciuria.

Other characteristics, aspects, objects and advantages of the invention will appear even more clearly on reading the description and the figures which follow, as well as various concrete examples, but in no way limiting, intended to illustrate it.

Description of the figures:
Figure 1: Effect of the association of 1,25-dihydroxyvitamin D3 and compound 1 (= 6 [3- (1-adamantyl) -4-hydroxyphenyl] -2-naphthanoic acid) on the inhibition of proliferation of keratinocytes. The results are expressed as a percentage of proliferation of keratinocytes relative to a control (DMSO) as a function of the concentration expressed in nM of 1,25-dihydroxyvitamin D3 in the absence (Ci) or in the presence () of compound 1 at a concentration 100 nM.

Figure 2: Effect of the association of 1,25-dihydroxyvitamin D3 and compound 1 on the inhibition of the proliferation of keratinocytes. The results are expressed as a percentage of proliferation of keratinocytes relative to a control (DMSO) as a function of the concentration expressed in nM of compound 1 in the presence of 1 nM of 1,25-dihydroxyvitamin D3 (Ci). In this figure, the value of compound 1 alone is also given at 100 nM (M).

6 [3- (1-adam antyl) -4-hydroxyphenyl] -2-naphtanoic acid and 6- [3- (1 adam antyl) 4-methoxyphenyl] -2-naphtanoic acid have already been described in the patent
US 4,717,720. Their derivatives can correspond to their esters, their amides, their salts, their alcohols or their aldehydes.

It is preferred to use 6 [3- (1-adamantyl) 4-hydroxyphenyl] -2-naphthanoic acid or its derivatives.

Among the ligands exhibiting activity for the VDR receptor, the following compounds may be mentioned:
- vitamin D3 (cholecalciferol),
- vitamin D2 (ergocalciferol),
- calcipotriol (or calcipotriene), sold in particular by the company LEO under the trade name DaivonexO,
- 25-hydroxyvitamin D3,
- 1a-hydroxyvitamin D3,
- 1 a, 25-dihydroxyvitamin D3 (calcitriol),
-1a, 25,26-trihydroxyvitamin D3,
-Ia, 23.25-trihydroxyvitamin D3,
- 24,25-dihydroxyvitamin D3,
- îa, 25-dihydroxyvitamin D2,
- 1a-hydroxyvitamin D2,
- îa, 24-dihydroxyvitamin D2.

 It is preferred to use 1 Ia, 25-dihydroxyvitamin D3.

In what follows, topical means any technique for administering a product by direct application of the latter to a superficial (or external) part of the body, such as the skin or the mucous membranes, and by systemic route, any technique for administering a product by a route other than topical, for example oral, enteral and / or parenteral.

The subject of the present invention is therefore, inter alia, a new pharmaceutical combination intended in particular for the treatment of the skin, characterized in that it comprises, in a physiologically acceptable support and compatible with the mode of administration chosen for this combination, at least one ligand having activity for the VDR receptor and at least one retinoid chosen from 6 [3- (1-adamantyl) 4-hydroxyphenyl] -2-naphthanoic acid, 6- [3- (1 - adam antyl) -4-m ethoxyphenyl) -2-naphthanoic or one of their derivatives as active ingredients.

The administration of the active agents (retinoid or ligand having activity for the VDR receptor) or of the combination according to the invention can be carried out by enteral, parenteral, topical or ocular route. However, preferably, the compositions (or combinations) according to the invention are packaged in a form suitable for topical application.

By the enteral route, the compositions may be in the form of tablets, capsules, dragees, syrups, suspensions, solutions, powders, granules, emulsions, microspheres or nanospheres or lipid or polymeric vesicles allowing controlled release. Parenterally, the compositions may be in the form of solutions or suspensions for infusion or for injection.

The mixtures of active ingredients in accordance with the invention are generally administered at daily doses of approximately 0.01 mg / kg to 100 mg / kg of body weight, and this at a rate of 1 to 3 doses / day.

By topical route, the pharmaceutical compositions, which are therefore more particularly intended for the treatment of the skin, can be in the form of ointments, creams, milks, ointments, powders, soaked tampons, solutions, gels. , sprays, lotions or suspensions. They can also be in the form of microspheres or nanospheres or lipid or polymeric vesicles or of polymeric patches and of hydrogels allowing a controlled release of the active agents. These compositions by topical route can moreover be presented either in anhydrous form or in an aqueous form. , according to the clinical indication.

The compositions for topical use in accordance with the invention contain the VDR ligand (s) at a concentration generally between 0.001% and 10% by weight, preferably between 0.1 and 1% by weight, relative to the total weight of the composition. Likewise, the compositions for topical use in accordance with the invention contain the retinoid or retinoids at a concentration generally between 0.001% and 10% by weight, preferably between 0.1 and 1% by weight, relative to the total weight of the composition.

By eye, these are mainly eye drops.

The compositions according to the invention can of course also contain inert or even pharmacodynamically active additives or combinations of these additives, and in particular: wetting agents; depigmenting agents such as hydroquinone, azelaic acid, caffeic acid or kojic acid; emollients; hydrating agents such as glycerol, PEG 400, thiamorpholinone, and its derivatives or even urea; antiseborrhoeic or anti-acne agents, such as S-carboxymethylcysteine, S benzyl-cysteamine, their salts or their derivatives, or benzoyl peroxide; antifungal agents such as ketoconazole or polymethylene 4,5 isothiazolidones-3; antibacterials, carotenoids and, in particular, - carotene; anti-psoriatic agents such as anthralin and its derivatives; and finally the eicosa-5,8,11,14-tetraynoic and eicosa-5,8,1 l-trynoic acids, their esters and amides.

The compositions according to the invention may also contain flavor-improving agents, preserving agents such as esters of parahydroxybenzoic acid, stabilizing agents, humidity-regulating agents, pH-regulating agents, agents osmotic pressure modifiers, emulsifiers, UV-A and UV-B filters, antioxidants, such as a4ocopherol, butylhydroxyanisole or butylhydroxytoluene.

The dermatological disorders linked to a hyperproliferation of the skin cells are in particular the following: 1) The dermatological affections linked to a keratinization disorder relating to cell proliferation in particular acne vulgaris, comedonian, polymorphic, rosacea, nodulocystic acne, conglobata , senile acne, secondary acne such as solar, medical or professional acne.

2) Other disorders of keratinization, in particular ichthyoses, ichthyosiform states, Darier's disease, palmoplantar keratoderma, leukoplakias and leukoplasiform states, cutaneous or mucous (buccal) lichen.

3) Other dermatological conditions linked to a keratinization disorder with an inflammatory and / or immuno-allergic component and, in particular, all forms of psoriasis, whether cutaneous, mucous or nail, and even psoriatic arthritis, or even cutaneous atopy, such as eczema or respiratory atopy or gingival hypertrophy.

4) All dermal or epidermal proliferations, whether benign or malignant, whether or not of viral origin such as common warts, flat warts and epidermodysplasia verruciformis, oral or florid papillomatosis and proliferations which may be induced by ultraviolet light, particularly in the case of basal and spinocellular epithelioma.

5) Other dermatological disorders, such as bullous dermatoses and collagen diseases.

6) Aging of the skin, whether photo-induced or chronological or for pigmentations and actinic keratoses, or any pathologies associated with chronological or actinic aging.

7) The stigma of epidermal and / or dermal atrophy induced by local or systemic corticosteroids, or any other form of skin atrophy.

8) Healing disorders or stretch marks.

9) Disorders of sebaceous function such as hyperseborrhea of acne or simple seborrhea.

10) Cancerous or precancerous conditions of the skin.

11) Any condition of viral origin in the skin.

12) Dermatological disorders with an immunological component.

The compositions according to the invention prove to be particularly effective for treatments 1) to 4), and more especially for the treatment of all forms of psoriasis.

We will now give, by way of non-limiting example, several examples intended on the one hand to demonstrate the effects attached to the present invention, and, on the other hand, to illustrate various concrete formulations in accordance with the invention.

EXAMPLE 1
The purpose of this example is to demonstrate the in vitro activity of the synergetic association in accordance with the invention on the proliferation of keratinocytes. The keratinocytes were obtained from skin specimen obtained from plastic surgery. They are used on the second pass.

The experimental protocol and the methods for determining proliferation were as follows:
Culture of keratinocytes:
The cells are cultured at 37 ° C. in a humid atmosphere in the presence of 5% CO 2 according to the Rheinwald and Green method in the presence of 3T3 fibroblates treated with mitomycin in MEM medium containing 10% (v / v) of serum. fetal calf (SVF), 0.4 pg / ml hydrocortisone, 10ng / ml EGF (Epidermal growth factor) and 10 M of chloleratoxin. 3T3 cells are
2 sown 24 hours before the keratinocytes at the rate of 15,000 cells per cm
Then, the keratinocytes are seeded at the rate of 4000 cells per
2 cm
Cell processing:
The cells are treated 2 hours after sowing the keratinocytes with compound 1 or 1,25-diOHvitamin D3 diluted in DMSO. The final concentration of DMSO in the culture medium does not exceed 0.2% (v / v). After 4 days of culture, the cells are harvested for the determination of proliferation.

Proliferation measurement:
The proliferation of the cells is determined by means of a kit referenced "Cell proliferation kit 11" (XTT assay) used according to the manufacturer's instructions (Boehringer, ref. 1465 05). It is expressed in absorbance units at 495 nm. The lower the absorbance measured, the more the proliferation of keratinocytes was inhibited.

The results obtained are collated in FIGS. 1 and 2. These figures quantify the change in the rate of proliferation of keratinocytes as a function of the concentration of the active agents used.

These figures clearly show that the combination of compound 1 and 1,25dihydroxyvitamin D3 has very good inhibition on the proliferation of keratinocytes.

 The same experiment was also carried out under identical conditions with a combination of 9-cis- or 13-cis-retinoic acid and 1,25dihydroxyvitamin D3. No inhibition synergy is observed on the proliferation of keratinocytes.

EXAMPLE 2
In this example, various concrete formulations have been illustrated based on associations in accordance with the invention (compound 1 is that defined above in the description).

A- ORAL ROUTE
(a) 0.2 g tablet
-Composel 0.001 g
- la, 25-dihydroxyvitamin D3 0.001 g
- Starch 0.113 9
- Dicalcium phosphate 0.020 g
- Silica 0.020 g
- Lactose 0.030 g
-Talc 0.010 g
- Magnesium stearate 0.005 g
(b) Oral suspension in 10 ml ampoules
- Compound 1 0.05 g
-1a, 25-dihydroxyvitamin D3 0.05 g
- Glycerin 1,0009
- Sorbitol at 70% 1,000g
- Sodium saccharinate 0.010 g
- Methyl parahydroxybenzoate 0.080 g
-Arome qs
- Purified water qs 10 ml
B- TOPICAL ROUTE
(a) Ointment - Compound 1 0.1 g -1 a, 25-dihydroxyvitamin D3 0.1 g - Isopropyl myristate 81.520 g - Fluid petroleum jelly 9.100 g - Silica ("Aerosil 200" sold by DEGUSSA) 9.180 g
(b) Non-ionic Water-in-Oil Cream - Compound 1 0.100 g -1a, 25-dihydroxyvitamin D3 0.100 g - Mixture of lanolin alcohol emulsifiers, waxes
and oils ("anhydrous Eucerin" sold by BDF) 39,900 g - Methyl parahydroxybenzoate 0.075 g - Propyl parahydroxybenzoate 0.075 g - Sterile demineralized water qs 100 g
(c) Lotion - Compound 1 0.100 g -1 a, 25-dihydroxyvitamin D3 0.100 g - Polyethylene glycol (PEG 400) 69.800 g - 95% ethanol 30,000 g
(d) Hydrophobic ointment - Compound 1 0.300 g -1 a, 25-dihydroxyvitamin D3 0.300 g - Isopropyl mirystate 36.400 g - Silicone oil ("Rhodorsil 47 V 300" sold
by RHONE-POULENC) 36,400 g - Beeswax 13,600 g - Silicone oil ("Abil 300,000 cst" sold
by GOLDSCHMIDT) qsp 1009
(e) Non-ionic Oil-in-Water Cream - Compound 1 0.500 g - îa, 25-dihydroxyvitamin D3 0.500 g - Cetyl alcohol 4,000 g - Glycerol monostearate 2,500 g - PEG stearate 50 2,500 g - Shea butter 9,200 g - Propylene glycol 2,000 g - Methyl parahydroxybenzoate 0.075 g - Propyl parahydroxybenzoate 0.075 g - Sterile demineralized water qs 100 g

Claims (13)

 CLAIMS 1- Association, characterized in that it comprises, in a physiologically acceptable support, at least one ligand having an activity for the VDR receptor and at least one retinoid chosen from 6 [3- (1-adamantyl) acid 4- hydroxyphenyl] -2-naphthanoic, 6- [3- (1-adamantyl) 4-methoxyphenyl] -2-naphtanoic acid or their derivatives. 2- Association according to claim 1 for use as a medicament. 3- Association according to one of claims 1 or 2, characterized in that the retinoid is chosen from 6 [3- (1-adamantyl) 4-hydroxyphenyl] -2-naphthanoic acid or its derivatives. 4- Association according to any one of the preceding claims, characterized in that the ligands having activity for the receptor VDR are chosen from: vitamin D3, vitamin D2, 25-hydroxyvitamin D3, la-hydroxyvitamin D3, la, 25-dihydroxyvitamin D3, la 1a, 25,26-trihydroxyvitamin D3, la 1a, 23,25-trihydroxyvitamin D3, la 24,25dihydroxyvitamin D3, la, 25-dihydroxyvitamin D2, la-hydroxyvitamin D2, la la, 24-dihydroxyvitamin D2. 5- Association according to the preceding claim, characterized in that the ligands having an activity for the VDR receptor is the ia, 25-dihydroxyvitamin D3. 6- Association according to any one of the preceding claims, characterized in that it is an association conditioned in a form suitable for enteral, parenteral, topical or ocular use. 7- Association according to the preceding claim, characterized in that it is an association conditioned in a form suitable for topical use. 8- Association according to any one of the preceding claims, characterized in that the ligand having an activity for nuclear receptors of VDR type is present at a concentration of between 0.001% and 10% by weight, preferably between 0.1 and 1% by weight, relative to the total weight of the association. 9- Association according to any one of the preceding claims, characterized in that the retinoid is present at a concentration of between 0.001% and 10% by weight, preferably between 0.1 and 1% by weight, relative to the weight total of the association. 10- Use of at least one retinoid chosen from 6 [3- (1-adamantyl) 4-hydroxyphenyl] -2-naphthanoic acid, 6- [3- (1-adam antyl) 4-m ethoxyphenyl acid ] -2naphtanoic and one of their derivatives for the manufacture of a pharmaceutical composition intended to increase the activity of inhibiting proliferation of skin cells, more particularly keratinocytes, due to at least one ligand having activity for the VDR receptor used for the treatment of dermatological disorders linked to hyperproliferation of skin cells. 11- Use of the combination according to any one of claims 1 to 9 for the manufacture of a dermatological composition intended to treat a dermatological disorder linked to a hyperproliferation of the skin cells, and more particularly of the keratinocytes. 12- Use according to one of claims 10 or 11, characterized in that the dermatological disorders linked to a hyperproliferation of the skin cells are chosen from the dermatological affections linked to a keratinization disorder relating to cell proliferation in particular acnes vulgar, comedonian, polymorphic, rosacea, nodulocystic acne, conglobata, senile acne, secondary acne such as solar, drug or professional acne, other keratinization disorders, in particular ichthyosis, ichthyosiform states, disease of Darier, palmoplantar keratoderma, leukoplakias and leukoplasiform states, cutaneous or mucous (oral) lichen, other dermatological conditions linked to a keratinization disorder with an inflammatory and / or immunoallergic component and, in particular, all forms of psoriasis whether it is cutaneous, mucous or nail, and even me psoriatic arthritis, or cutaneous atopy, such as eczema or respiratory atony or gingival hypertrophy, all dermal or epidermal proliferations whether benign or malignant, whether or not d 'viral origin such as common warts, flat warts and epidermodysplasia verruciformis, oral or florid papillomatosis and the proliferations which can be induced by ultraviolet rays, in particular in the case of baso and spinocellular epithelioma. 13- Use according to the preceding claim, characterized in that the dermatological disorders linked to a hyperproliferation of the skin cells are all forms of psoriasis.